EP1636266A2 - Polypeptides de fusion de variants de glycoproteines ib alpha de plaquette et leurs methodes - Google Patents
Polypeptides de fusion de variants de glycoproteines ib alpha de plaquette et leurs methodesInfo
- Publication number
- EP1636266A2 EP1636266A2 EP04776599A EP04776599A EP1636266A2 EP 1636266 A2 EP1636266 A2 EP 1636266A2 EP 04776599 A EP04776599 A EP 04776599A EP 04776599 A EP04776599 A EP 04776599A EP 1636266 A2 EP1636266 A2 EP 1636266A2
- Authority
- EP
- European Patent Office
- Prior art keywords
- polypeptide
- amino acid
- seq
- acid sequence
- protein
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/745—Blood coagulation or fibrinolysis factors
- C07K14/755—Factors VIII, e.g. factor VIII C (AHF), factor VIII Ag (VWF)
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/62—DNA sequences coding for fusion proteins
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
Definitions
- vascular inflammation and repair response involves adhesive interactions between various cell types normally found freely circulating in blood. Examples of such interactions include those that can occur between platelets, leukocytes and the inner wall of blood vessels (i.e., the vascular endothelium). Under conditions of high fluid shear forces, platelets adhere to the endothelium via an interaction between the glycoprotein (GP) Ib-LX-V complex on their surface and von Willebrand factor (vWF) present on exposed vessel subendothelium.
- GP glycoprotein
- Ib-LX-V complex the inner wall of blood vessels
- vWF von Willebrand factor
- platelets adhering to the vascular endothelium can bind and capture freely circulating platelets via vWF-mediated tethering, enabling thrombus growth through successive layers of platelets.
- the GPIboc chain of the GPIb-LX-V complex can also facilitate the binding of ⁇ -thrombin to the platelet surface, enhancing the thromb ⁇ n-mediated cleavage of GPV and protease-activated receptors (PARs).
- PARs protease-activated receptors
- leukocytes can adhere to activated endothelium either directly, or indirectly by first adhering to vWF-immobilized platelets.
- leukocyte cell surface molecules that bind to either the selectin or integrin classes of adhesion receptors mediate these adhesion events.
- Leukocyte-platelet adhesion is thought to occur, in part, via interaction of the leukocyte surface integrin molecule Mad and the GPlb component of the platelet surface GPIb-LX-V complex.
- vascular disturbances such as atherosclerotic plaque rupture or mechanical injury, e.g., such as that caused by angioplasty, stent placement, cardiopulmonary bypass procedures, ischemic damage or stenosis
- leukocytes and platelets can accumulate at a vascular lesion site and provide multiple adhesive substrates for one another. This accumulation of leukocytes and platelets leads to the local production of factors including, e.g., mitogens, cytokines and chemokines, causing the further undesirable progression of a vascular disease.
- Therapeutic polypeptides including a v F-binding region derived from GPIb ⁇ have been described.
- One such polypeptide is based on a sequence containing two amino acid substitutions (G233V M239V) in the amino acid sequence of wild-type human GPIb ⁇ .
- the invention provides improved glycoprotein-Iboc variant polypeptides that are useful as therapeutic agents for treating vascular disorders.
- the variants show decreased aggregation, enhanced stability, decreased bindmg to thrombin, or two or more of these properties.
- One use for the glycoprotein-Ib ⁇ -protein variant polypeptides is as a fusion protein to treat vascular conditions associated with vascular inflammation, thrombosis, atherosclerosis, and angioplasty-related restenosis.
- the polypeptide binds with lower affinity to alpha thrombin relative to binding to alpha thrombin of a polypeptide that includes the naturally occurring amino acid sequence of human GPIb ⁇ (SEQ ID NO: 1) or a polypeptide, termed GPIb2V (SEQ ID NO:2), which includes the amino acid substitutions G233V and M239V relative to the amino acid sequence of SEQ ID NO:l.
- GPIb2V SEQ ID NO:2V
- polypeptides that show reduced thrombin binding are polypeptides that include one, two, or three of the amino acid substitutions Y276F, Y278F, Y279V, or a conservative variant thereof, relative to the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO:2.
- aggregation of the polypeptide is lowered relative to aggregation of a polypeptide that includes the amino acid sequence of human SEQ ID NO:l or SEQ ID NO:2.
- decreased aggregation is observed during synthesis of the polypeptide in a cell.
- An example of a polypeptide that shows decreased aggregation is a polypeptide that includes the amino acid substitution C65S, or a conservative variant thereof, relative to the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO:2.
- the polypeptide is more resistant to proteolysis than a polypeptide that includes the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO:2.
- An example of such a polypeptide is a polypeptide includes the amino acid substitution K237V, or a conservative variant thereof, relative to the amino acid sequence of SEQ ID NO:l or SEQ ID NO:2.
- polypeptides examples include those that have the following amino acid sequence substitutions relative to the amino acid sequence of SEQ ID NO:l or SEQ ID NO:2:Y276F, Y276F K237V,Y276F C65S, Y276F Y278F Y279F, Y276F Y278F Y279F K237V and Y276F Y278F Y279F K237V C65S.
- the polypeptides of the invention can be provided as fusion proteins.
- the glycoprotein Ib ⁇ fusion protein can include a first polypeptide, comprising at least a region of a glycoprotein Ib ⁇ polypeptide variant, linked to a second polypeptide.
- the second polypeptide forms a multimer, e.g., a dimer.
- the second polypeptide includes at least a region of an immunoglobulin polypeptide.
- nucleic acid encoding the glycoprotein-Ib ⁇ variant polypeptide, as well as vectors, cells and methods of expressing the glycoprotein-Ib ⁇ variant polypeptide using glycoprotein-Ib ⁇ variant polypeptide-encoding nucleic acids.
- the invention additionally includes a nucleic acid encoding a glycoprotein Ib ⁇ variant fusion polypeptide, as well as a vector containing glycoprotein Ib ⁇ fusion polypeptide-encoding nucleic acids described herein, and a cell containing the vectors or nucleic acids described herein.
- Also provided by the invention is a method of inhibiting leukocyte adhesion to a biological tissue by contacting a leukocyte with a glycoprotein Ib ⁇ fusion polypeptide.
- the leukocyte is contacted in an amount sufficient to inhibit adherence of the leukocyte and the biological tissue
- the invention provides a method of treating a disorder associated with platelet activation.
- the method includes administering to a subject an effective amount of a glycoprotein Ib ⁇ fusion polypeptide.
- pharmaceutical compositions that include glycoprotein Ib ⁇ variant polypeptides, or glycoprotein Ib ⁇ variant fusion polypeptides.
- FIG. 1 is a graph showing inhibition of GPIba chimeras on ristocetin-induced human platelet aggregation. Shown is light transmittance (y-axis) for wild-type GPIba ("Normal"), and GPIb-Ig WT, GPIba-290/2V-Ig, GPIba-290/2V/FFF-Ig chimeras.
- FIG. 2 is a graph showing the antithrombotic efficacy of GPIb ⁇ chimeras in a canine Folts' model of coronary thrombosis (y-axis: LCX blood).
- FIG. 3 is a graph showing inhibition of platelet function by GPIb/290/2V-Ig and GPIb/290/2V/FFF-Ig in a canine model of coronary thrombosis assessed by PFA-100.
- Glycoprotein Ib ⁇ -derived polypeptides with decreased binding to alpha thrombin, decreased aggregation and/or increased resistance to proteolysis according to the invention are useful as therapeutic agents in, e.g., treating various conditions that benefit by inhibiting bindmg of activated platelet cells to vWF on vascular cells.
- the polypeptides are provided as fusion proteins.
- the protein variants of the invention with decreased thrombin binding advantageously result in decreased bleeding. Binding of thrombin to the platelet GPIb ⁇ receptor is necessary for clotting. Thrombin binding to a therapeutic soluble GPIb can augment bleeding in vivo by sequestering thrombin away from the platelet GPIb ⁇ receptor, thus reducing thrombin-induced platelet aggregation. Thus, the protein variants that show lower affinity for thrombin make the thrombin available to interact with the platelet IB ⁇ receptor, which in turn promotes clotting.
- a suitable polypeptide includes an amino acid sequence with one to 15 amino acid substitutions, deletions or insertions relative to amino acids in the region between, and including, amino acids 65-279 of a naturally occurring human GPIb ⁇ protein sequence shown in SEQ ID NO:l, below, or of the variant GPIb2V, whose amino acid sequence is shown in SEQ ID NO:2, below.
- the polypeptide has one or more of the following activities: (i) lower affinity binding to alpha thrombin relative to binding to alpha thrombin of a polypeptide that includes the amino acid sequence of human SEQ ID NO:l or SEQ ID NO:2; (ii) lower aggregation relative to aggregation of a polypeptide that includes the amino acid sequence of SEQ ID NO:l or SEQ ID NO:2; or (iii) increased resistance to proteolysis relative to a polypeptide that includes the amino acid sequence of SEQ ID NO:l or SEQ ID NO:2.
- amino acid sequence of the GPIb2Vvariant is provided below as SEQ ID NO:2. This variant is also discussed in US patent application publication number 20030091576, and its counterpart WO 02/063003.
- the polypeptide has no more than 12 substitutions, insertions, or deletions relative to the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO:2. For example, it may have 10, 8, 7, 6, 5 or fewer substitutions, insertions, or deletions relative to the amino acid sequence of SEQ ID NO:l or SEQ ID NO:2. In some embodiments, the substitutions, insertions, or deletions are in the region between amino acids 65 and 279, inclusive.
- the polypeptide binds with lower affinity to alpha thrombin relative to binding to alpha thrombin of a polypeptide that includes the amino acid sequence of SEQ ID NO:l or SEQ ID NO:2.
- polypeptides that show reduced binding are polypeptides that include one, two, or three of the amino acid substitutions Y276F, Y278F, Y279V, or a conservative variant thereof, relative to the amino acid sequence of SEQ ID NO:l or SEQ ID NO:2.
- aggregation of the polypeptide is lowered relative to aggregation of a polypeptide that includes the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO:2.
- decreased aggregation is observed during synthesis of the polypeptide in a cell.
- An example of a polypeptide that shows decreased aggregation is a polypeptide that includes the amino acid substitution C65S, or a conservative variant thereof, relative to the amino acid sequence of SEQ ID NO:l or SEQ ID NO:2.
- the polypeptide is more resistant to proteolysis than a polypeptide that includes the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO:2.
- polypeptide includes the amino acid substitution K237V, or a conservative variant thereof, relative to the amino acid sequence of SEQ ID NO:l or SEQ ID NO:2.
- the polypeptide includes an amino acid sequence with one to 10 amino acid substitutions, insertions, or deletions relative to amino acids 1-290 of SEQ ID NO:l or SEQ ID NO:2, provided that at least one of the amino acid substitutions is C65S, K237V,Y276F Y278F, Y279F, K237V.
- polypeptides examples include those that have the following amino acid sequence substitutions relative to the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO:2: Y276F, Y276F K237V,Y276F C65S, Y276F Y278F Y279F, Y276F Y278F Y279F K237V and Y276F Y278F Y279F K237V C65S.
- the invention also includes polypeptides with one or more, e.g., 2, 3, 5, 6, 8, 10, 15 or more amino acid substitutions in polypeptides derived from glycoprotein IB ⁇ sequences in addition to those corresponding to SEQ ID NO: 1 and SEQ ID NO:2.
- polypeptides with one or more, e.g., 2, 3, 5, 6, 8, 10, 15 or more amino acid substitutions in polypeptides derived from glycoprotein IB ⁇ sequences in addition to those corresponding to SEQ ID NO: 1 and SEQ ID NO:2.
- GPIb302 SEQ ID NO:3
- GPIb302/2A SEQ ID NO:4
- GPIb/4X SEQ ID NO:5
- GBIb290/l A SEQ ID NO:6
- a glycoprotein protein variant can be provided as part of a fusion protein.
- glycoprotein Ib ⁇ protein-immunoglobulin fusion proteins are useful for inhibiting adherence of platelets and leukocytes to biological tissues, such as for example the vascular endothelium.
- the fusion proteins of the invention, or nucleic acids encoding these fusion proteins can be incorporated into pharmaceutical compositions and administered to a subject to inhibit an interaction between an glycoprotein Ib ⁇ ligand (such as von Willebrand Factor, Mac-1, P-selectin or thrombin) and an glycoprotein Ib ⁇ protein on the surface of a cell, such as a platelet.
- an glycoprotein Ib ⁇ ligand such as von Willebrand Factor, Mac-1, P-selectin or thrombin
- glycoprotein Ib ⁇ protein-mediated platelet aggregation and associated signal transduction suppresses glycoprotein Ib ⁇ protein -mediated platelet aggregation and associated signal transduction in vivo.
- the glycoprotein Ib ⁇ protein-immunoglobulin fusion proteins can be used to modulate the bioavailability of a glycoprotein Ib ⁇ protein cognate ligand.
- Inhibition of the glycoprotein Ib ⁇ protein ligand/glycoprotein Ib ⁇ protein interaction are useful therapeutically for, inter alia, the treatment of vascular inflammation and other vascular disorders associated with platelet activation.
- Glycoprotein Variant Ib ⁇ Fusion Polypeptides in various aspects provides fusion proteins that include a first polypeptide containing at least a portion of a glycoprotein Ib ⁇ polypeptide variant operatively linked to a second polypeptide.
- a glycoprotein Ib ⁇ "fusion protein” or “chimeric protein” includes at least a portion of a glycoprotein Ib ⁇ polypeptide variant operatively linked to a non-glycoprotein Ib ⁇ polypeptide.
- glycoprotein Ib ⁇ polypeptide or “glycoprotein Ib ⁇ polypeptide variant” refers to a polypeptide having an amino acid sequence corresponding to at least a portion of a glycoprotein Ib ⁇ polypeptide
- a “non-glycoprotein Ib ⁇ polypeptide” refers to a polypeptide having an amino acid sequence corresponding to a protein that is not substantially homologous to the glycoprotein Ib ⁇ protein, e.g., a protein that is different from the glycoprotein Ib ⁇ polypeptide or fragment and that is derived from the same or a different organism.
- the glycoprotein Ib ⁇ polypeptide can correspond to all or a portion of an Ib ⁇ protein.
- a glycoprotein Ib ⁇ fusion protein comprises at least one biologically active portion of a glycoprotein Ib ⁇ protein. In another embodiment, a glycoprotein Ib ⁇ fusion protein comprises at least two biologically active portions of a glycoprotein Ib ⁇ protein. In yet another embodiment, a glycoprotein Ib ⁇ fusion protein comprises at least three biologically active portions of a glycoprotein Ib ⁇ protein.
- the term "operatively linked" is intended to indicate that the first and second polypeptides are linked in a manner that allows for at least one function associated with a glycoprotein Ib ⁇ polypeptide.
- the term operatively linked means that a nucleic acid encoding the glycoprotein Ib ⁇ polypeptide and the non-glycoprotein Ib ⁇ polypeptide are fused in-frame to each other.
- the non-glycoprotein Ib ⁇ polypeptide can be fused to the N-terminus or C-terminus of the glycoprotein Ib ⁇ polypeptide.
- the glycoprotein Ib ⁇ fusion protein may be linked to one or more additional moieties.
- the glycoprotein Ib ⁇ fusion protein may additionally be linked to a GST fusion protein in which the glycoprotein Ib ⁇ fusion protein sequences are fused to the C-terminus of the GST (i.e., glutathione S-transferase) sequences.
- GST glutathione S-transferase
- the fusion protein includes a heterologous signal sequence (i.e., a polypeptide sequence that is not present in a polypeptide encoded by a glycoprotein Ib ⁇ nucleic acid) at its N-terminus.
- the native glycoprotein Ib ⁇ signal sequence can be removed and replaced with a signal sequence from another protein.
- glycoprotein Ib ⁇ In certain host cells (e.g., mammalian host cells), expression and/or secretion of glycoprotein Ib ⁇ can be increased through use of a heterologous signal sequence.
- a representative signal sequence is MPLLLLLLLLPSPLFIP (SEQ ID NO:8).
- Another representative signal sequence is MPLQLLLLLI LLGPGNSLQL WDTWADEAEK ALGPLLARDRR . (SEQ ID NO:9).
- one or more amino acids can additionally be inserted between the first polypeptide moiety comprising the GP Ib ⁇ moiety and the second polypeptide moiety.
- An chimeric or fusion protein of the invention can be produced by standard recombinant DNA techniques.
- DNA fragments coding for the polypeptide variant sequences are ligated together in-frame in accordance with conventional techniques, e.g., by employing blunt-ended or stagger-ended termini for ligation, restriction enzyme digestion to provide for appropriate termini, filling-in of cohesive ends as appropriate, alkaline phosphatase treatment to avoid undesirable joining, and enzymatic ligation.
- the fusion gene can be synthesized by conventional techniques including automated DNA synthesizers.
- PCR amplification of gene fragments can be carried out using anchor primers that give rise to complementary overhangs between two consecutive gene fragments that can subsequently be annealed and reamplified to generate a chimeric gene sequence (see, for example, Ausubel et al. (eds.) CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, 1992).
- anchor primers that give rise to complementary overhangs between two consecutive gene fragments that can subsequently be annealed and reamplified to generate a chimeric gene sequence
- a fusion moiety e.g., an Fc region of an immunoglobulin heavy chain.
- a glycoprotein Ib ⁇ encoding nucleic acid can be cloned into such an expression vector such that the fusion moiety is linked in-frame to the immunoglobulin protein.
- Glycoprotein Ib ⁇ fusion polypeptides may exist as oligomers, such as dimers or trimers.
- the glycoprotein Ib ⁇ fusion polypeptide is a dimer.
- the GP lb ⁇ polypeptide moiety is provided as a variant GP lb ⁇ polypeptide having a mutation in the naturally-occurring GP lb ⁇ sequence (wild type) that results in higher affinity (relative to the non-mutated sequence) binding of the GP I ⁇ polypeptide to a leukocyte cell surface molecule.
- the mutant polypeptide may bind with higher affinity to von Willebrand factor (vWF). This increased reactivity, or hyperresponsiveness, can be assessed using low concentrations of ristocetin.
- vWF von Willebrand factor
- any other suitable means for determining the reactivity of the polypeptide with vWF can also be utilized to identify polypeptides which are "more" reactive with vWF, i.e. more reactive than naturally-occurring wild-type GP Ib ⁇ .
- Examples of GP lb ⁇ polypeptide variants that bind with higher affinity to vWF include GP Ib ⁇ variants that include sequence alterations in the hinge region of a GP Ib ⁇ polypeptide. The hinge region is defined as the region including residues 220 to 310 and is reported to be a major binding site for vWF within the GP lb ⁇ polypeptide.
- Mutations in the hinge region include those at residue 233, which in the wild-type GP lb ⁇ encodes glycine.
- An example of a suitable substitution of is one in which the glycine at position 233 is replaced with valine (i.e., G233V).
- a second site for mutation at the hinge region is at residue 239, which in the wild-type GP lb ⁇ encodes methionine.
- a substitution of valine for methionine 239 is a representative substitution, but other amino acids can also be substituted.
- hinge region variants of GP lb ⁇ polypeptides suitable for use in a fusion polypeptide of the invention have mutations at residue both positions 233 and 239.
- the invention includes fusion proteins that have a substitution at position 239, e.g., an M239V substitution of a variant GP lb ⁇ polypeptide.
- a fusion protein having a substitution at position 233 e.g., G233V
- the first polypeptide includes a full-length GP lb ⁇ polypeptide.
- the first polypeptide comprise less than full-length GP lb ⁇ polypeptide.
- the first polypeptide less than 600 amino acids in length, e.g., less than or equal to 500, 250, 150, 100, 50, or 25 amino acids in length.
- the second polypeptide is soluble. In some embodiments, the second polypeptide enhances the half-life, (e.g., the serum half-life) of the linked polypeptide. In some embodiments, the second polypeptide includes a sequence that facilitates association of the fusion polypeptide with a second GP lb ⁇ polypeptide. In some embodiments, the second polypeptide includes at least a region of an immunoglobulin polypeptide. Immunoglobulin fusion polypeptide are known in the art and are described in e.g., US Patent Nos. 5,516,964; 5,225,538; 5,428,130;5,514,582; 5,714,147;and 5,455,165.
- the second polypeptide comprises a full-length immunoglobulin polypeptide.
- the second polypeptide comprise less than full-length immunoglobulin polypeptide, e.g., a heavy chain, light chain, Fab, Fab 2 , Fv, or Fc.
- the second polypeptide includes the heavy chain of an immunoglobulin polypeptide.
- the second polypeptide includes the Fc region of an immunoglobulin polypeptide.
- the second polypeptide has less effector function that the effector function of a Fc region of a wild-type immunoglobulin heavy chain.
- Fc effector function includes for example, Fc receptor binding, complement fixation and T cell depleting activity (see, or example, US Patent No. 6,136,310). Methods of assaying T cell depleting activity, Fc effector function, and antibody stability are known in the art.
- the second polypeptide has low or no affinity for the Fc receptor. In an alternative embodiment, the second polypeptide has low or no affinity for complement protein Clq.
- a representative second polypeptide sequence includes the amino acid sequence of SEQ ID NO: 10. This sequence includes a Fc region. Underlined amino acids are those that differ from the amino acid found in the corresponding position of the wild-type immunoglobulin sequence:
- HTCPPCPAPEALGAPSVFLFPPKPKDTL ISRTPEVTCVWDVSHEDPEVKFN YVDGVEVHNAKTKPREEQ YNSTYRVVSVLTVLHQD LNGKEYKCKVSNKALPVPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTC LVKGFYPSDIAVEWESNGQPENNYKTTPPV DSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQK S SLSPGK (SEQ ID NO: 10)
- the GPIb ⁇ polypeptide variants can include, in addition to the designated substituted amino acid substitution, an alternative amino acid that fulfills the same function as the designated acid.
- the alternative amino acid is related to the designated amino acid as a conservative amino acid substitution of the indicated amino acid.
- a "conservative amino acid substitution” is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains ⁇ e.g.
- lysine, arginine, histidine acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g , threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine).
- acidic side chains e.g., aspartic acid, glutamic acid
- uncharged polar side chains e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine
- the tyrosine residue can be replaced with tryptophan or histidine.
- the cysteine may alternatively be replaced with glycine, asparagine, glutamine, threonine, or tyrosine.
- the lysine may can alternatively be replaced with alanine, leucine, isoleucine, proline, phenylalanine, methionine, or tryptophan.
- the invention also includes a polypeptide whose sequence that are 80%, 85%, 90%, 95%, 98%, or 99% homologous to the sequence disclosed in SEQ ID NO:l or SEQ ID NO:2, provided that it includes one or more of the amino acid substations C65S, K237V,Y276F Y278F, Y279F, K237V.
- nucleic acids or proteins of the invention include, but are not limited to, molecules comprising regions that are substantially homologous to the nucleic acids or proteins of the invention, in various embodiments, by at least about 70%, 80%, 85%, 90%, 95%, 98%, or even 99% identity (e.g., an identity of 80-99%) over a nucleic acid or amino acid sequence of identical size or when compared to an aligned sequence in which the alignment is done by a computer homology program known in the art, or whose encoding nucleic acid is capable of hybridizing to the complement of a sequence encoding the aforementioned proteins under stringent, moderately stringent, or low stringent conditions.
- An exemplary program is the Gap program (Wisconsin Sequence Analysis Package, Version 8 for UN X, Genetics Computer Group, University Research Park, Madison, WI) using the default settings, which uses the algorithm of Smith and Waterman.
- Another aspect of the invention pertains to vectors (including expression vectors) containing a nucleic acid encoding glycoprotein Ib ⁇ fusion polypeptides, or derivatives, fragments, analogs or homologs thereof.
- Recombinant expression vectors of the invention can be designed for expression of glycoprotein Ib ⁇ fusion polypeptides in prokaryotic or eukaryotic cells.
- the glycoprotein Ib ⁇ fusion polypeptide expression vector is a yeast expression vector.
- yeast Saccharomyces cerevisiae examples of vectors for expression in yeast Saccharomyces cerevisiae are known in the art.
- glycoprotein Ib ⁇ fusion polypeptide can be expressed in insect cells using methods known in the art, e.g., with baculovirus expression vectors.
- a nucleic acid of the invention is expressed in mammalian cells using a mammalian expression vector.
- the recombinant mammalian expression vector is capable of directing expression of the nucleic acid preferentially in a particular cell type (e.g., tissue-specific regulatory elements are used to express the nucleic acid). Tissue-specific regulatory elements are known in the art.
- the invention further provides a recombinant expression vector comprising a DNA molecule of the invention cloned into the expression vector in an antisense orientation. That is, the DNA molecule is operatively-linked to a regulatory sequence in a manner that allows for expression (by transcription of the DNA molecule) of an RNA molecule that is antisense to a glycoprotein Ib ⁇ fusion polypeptide rnRNA.
- Regulatory sequences operatively linked to a nucleic acid cloned in the antisense orientation can be chosen that direct the continuous expression of the antisense RNA molecule in a variety of cell types, for instance viral promoters and/or enhancers, or regulatory sequences can be chosen that direct constitutive, tissue specific or cell type specific expression of antisense RNA.
- the antisense expression vector can be in the form of a recombinant plasmid, phagemid or attenuated virus in which antisense nucleic acids are produced under the control of a high efficiency regulatory region, the activity of which can be determined by the cell type into which the vector is introduced.
- a host cell can be any prokaryotic or eukaryotic cell.
- glycoprotein Ib ⁇ fusion polypeptides can be expressed in bacterial cells such as E. coli, insect cells, yeast or mammalian cells (such as human, Chinese hamster ovary cells (CHO) or COS cells).
- bacterial cells such as E. coli, insect cells, yeast or mammalian cells (such as human, Chinese hamster ovary cells (CHO) or COS cells).
- mammalian cells such as human, Chinese hamster ovary cells (CHO) or COS cells.
- Other suitable host cells are known to those skilled in the art.
- Vector DNA can be introduced into prokaryotic or eukaryotic cells via conventional transformation or transfection techniques.
- Mammalian host cell such as a Chinese hamster ovary cells (CHO) or COS cells can be transfected with expression vectors to enable, via posttranslational modification, the generation of the sialyl Lewis epitope on the N-linked and O-linked glycans of glycoprotein Ib ⁇ fusion polypeptides.
- CHO cells this requires the co-expression of an ⁇ - 1,3/1, 4 fucosyltranseferase (Kukowska-Latallo et al., Genes Dev. 4: 1288-303, 1990 ) and Core2 beta-1 ,6-N- acetylglucosaminyltransferase enzymes.
- a gene that encodes a selectable marker (e.g., resistance to antibiotics) is generally introduced into the host cells along with the gene of interest.
- selectable markers include those that confer resistance to drugs, such as G418, hygromycin and methotrexate.
- Nucleic acid encoding a selectable marker can be introduced into a host cell on the same vector as that encoding glycoprotein Ib ⁇ fusion polypeptides or can be introduced on a separate vector. Cells stably transfected with the introduced nucleic acid can be identified by drug selection (e.g., cells that have incorporated the selectable marker gene will survive, while the other cells die).
- a host cell of the invention such as a prokaryotic or eukaryotic host cell in culture, can be used to produce (i.e., express) glycoprotein Ib ⁇ fusion polypeptides.
- the invention further provides methods for producing glycoprotein Ib ⁇ fusion polypeptides using the host cells of the invention.
- the method comprises culturing the host cell of invention (into which a recombinant expression vector encoding glycoprotein Ib ⁇ fusion polypeptides has been introduced) in a suitable medium such that glycoprotein Ib ⁇ fusion polypeptides is produced.
- the method further comprises isolating glycoprotein Ib ⁇ fusion polypeptide from the medium or the host cell.
- the fusion polypeptides may be isolated and purified in accordance with conventional conditions, such as extraction, precipitation, chromatography, affinity chromatography, electrophoresis or the like. Chemical synthesis of polypeptides facilitates the incorporation of modified or unnatural amino acids, including D-amino acids and other small organic molecules. Replacement of one or more L-amino acids in a peptide with the corresponding D-amino acid isoforms can be used to increase the resistance of peptides to enzymatic hydrolysis, and to enhance one or more properties of biologically active peptides, i.e., receptor binding, functional potency or duration of action.
- Macrocyclization is often accomplished by forming an amide bond between the peptide N- and C-termini, between a side chain and the N- or C-terminus [e.g., with K 3 Fe(CN) 6 at pH 8.5], or between two amino acid side chains.
- Disulfide bridges are also introduced into linear sequences to reduce their flexibility.
- the replacement of cysteine residues with penicillamine (Pen, 3-mercapto-(D) valine) has been used to increase the selectivity of some opioid-receptor interactions.
- compositions Including Glycoprotein Ib ⁇ Fusion Polypeptides or Nucleic Acids Encoding Same
- glycoprotein Ib ⁇ fusion proteins or nucleic acid molecules encoding these fusion proteins, (also referred to herein as "Therapeutics” or “active compounds”) of the invention, and derivatives, fragments, analogs and homologs thereof, can be incorporated into pharmaceutical compositions suitable for administration.
- Such compositions typically comprise the nucleic acid molecule, protein, or antibody and a pharmaceutically acceptable carrier.
- pharmaceutically acceptable carrier is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration.
- Suitable carriers are described in the most recent edition of Remington's Pharmaceutical Sciences, a standard reference text in the field, which is incorporated herein by reference.
- Examples of such carriers or diluents include, but are not limited to, water, saline, finger's solutions, dextrose solution, and 5% human serum albumin. Liposomes and non-aqueous vehicles such as fixed oils may also be used.
- the use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, use thereof in the compositions is contemplated. Supplementary active compounds can also be incorporated into the compositions.
- the active agents disclosed herein can also be formulated as liposomes.
- Liposomes are prepared by methods known in the art. Liposomes with enhanced circulation time are disclosed in U.S. PatentNo. 5,013,556.
- Particularly useful liposomes can be generated by the reverse-phase evaporation method with a lipid composition comprising phosphatidylcholine, cholesterol, and PEG-derivatized phosphatidylethanolamine (PEG-PE). Liposomes are extruded through filters of defined pore size to yield liposomes with the desired diameter.
- a pharmaceutical composition of the invention is formulated to be compatible with its intended route of administration. Examples of routes of administration include parenteral, e.g., intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (i.e., topical), transmucosal, and rectal administration.
- Solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid (EDTA); buffers such as acetates, citrates or phosphates, and agents for the adjustment of tonicity such as sodium chloride or dextrose.
- the pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide.
- the parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.
- compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion.
- suitable carriers include physiological saline, bacteriostatic water, Cremophor EL (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS).
- the composition must be sterile and should be fluid to the extent that easy syringeability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi.
- the carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof.
- the proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.
- Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like.
- isotonic agents can be included in the composition, for example, sugars, polyalcohols such as manitol and sorbitol; and sodium chloride.
- Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate and gelatin.
- Sterile injectable solutions can be prepared by incorporating the active compound (e.g., a glycoprotein Ib ⁇ fusion protein) in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization.
- dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above.
- methods of preparation are vacuum drying and freeze-drying that yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.
- Oral compositions generally include an inert diluent or an edible carrier. They can be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash, wherein the compound in the fluid carrier is applied orally and swished and expectorated or swallowed. Pharmaceutically compatible binding agents, and/or adjuvant materials can be included as part of the composition.
- the tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.
- a binder such as microcrystalline cellulose, gum tragacanth or gelatin
- an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch
- a lubricant such as magnesium stearate or Sterotes
- a glidant such as colloidal silicon dioxide
- the compounds are delivered in the form of an aerosol spray from pressured container or dispenser which contains a suitable propellant, e.g., a gas such as carbon dioxide, or a nebulizer.
- a suitable propellant e.g., a gas such as carbon dioxide, or a nebulizer.
- Systemic administration can also be by transmucosal or transdermal means.
- penetrants appropriate to the barrier to be permeated are used in the formulation.
- penetrants are generally known in the art, and include, for example, for transmucosal administration, detergents, bile salts, and fusidic acid derivatives.
- Transmucosal administration can be accomplished through the use of nasal sprays or suppositories.
- the active compounds are formulated into ointments, salves, gels, or creams as generally known in the art.
- the compounds can also be prepared in the form of suppositories (e.g., with conventional suppository bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.
- the active compounds are prepared with carriers that will protect the compound against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems.
- Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparation of such formulations will be apparent to those skilled in the art.
- the materials can also be obtained commercially from Alza Corporation and Nova Pharmaceuticals, Inc.
- Liposomal suspensions can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art.
- oral or parenteral compositions are formulated in dosage unit form for ease of administration and uniformity of dosage.
- Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subject to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier.
- the specification for the dosage unit forms of the invention are dictated by and directly dependent on the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and the limitations inherent in the art of compounding such an active compound for the treatment of individuals.
- the nucleic acid molecules of the invention can be inserted into vectors and used as gene therapy vectors.
- Gene therapy vectors can be delivered to a subject by, for example, intravenous injection, local administration or by stereotactic injection.
- the pharmaceutical preparation of the gene therapy vector can include the gene therapy vector in an acceptable diluent, or can comprise a slow release matrix in which the gene delivery vehicle is imbedded.
- the pharmaceutical preparation can include one or more cells that produce the gene delivery system. Sustained-release preparations can be prepared, if desired.
- sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the antibody, which matrices are in the form of shaped articles, e.g., films, or microcapsules.
- sustained-release matrices include polyesters, hydrogels (for example, poly(2-hydroxyethyl- methacrylate), or poly(vinylalcohol)), polylactides , copolymers of L-glutamic acid and ⁇ ethyl-L- glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as the LUPRON DEPOT TM (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), and poly-D-(-)-3-hydroxybutyric acid.
- polyesters for example, poly(2-hydroxyethyl- methacrylate), or poly(vinylalcohol)
- compositions can be included in a container, pack, or dispenser together with instructions for administration.
- Included in the invention is a method of inhibiting adherence of a blood cell to a biological tissue in a biological system.
- the method includes adding to a biological system a fusion polypeptide of the invention in an amount sufficient to inhibit adherence of a blood cell to the biological tissue.
- the blood cell can be for example, a leukocyte, platelet or red blood cell.
- the leukocyte can be any leukocyte that is capable of adhering to a biological tissue.
- the leukocyte is a granulocyte, (i.e., neutrophil, basophil or eosinohil), monocyte (i.e., macrophage) or lymphocyte (e.g., T-lymphocyte, B-lymphocyte, tumor infiltrating lymphocytes or natural killer cell).
- the leukocytes express a ⁇ 2 intergin, e.g. Mac-1.
- the leuckocyte expresses a selectin ligand.
- Also included in the inventions are methods of inhibiting adherence of a protein to a biological tissue in a biological system.
- the method includes adding to a biological system a fusion polypeptide of the invention in an amount sufficient to inhibit adherence of the protein to the biological tissue.
- the protein can be membrane associated (e.g., covalently, non- covalently, or ionic). Alternatively, the protein can be in a soluble form (i.e., in solution).
- the protein is von Willebrand Factor, tlirombin, P-selectin of glycoprotein Ib ⁇ .
- a biological system is meant to include any system that comprises biological components, e.g., cells, proteins, carbohydrates, lipids or nucleic acids.
- the biological system can be an in vivo, ex vivo or in vitro system.
- “Adherence" is meant to include any biological interaction of a leukocyte, e.g., rolling, firm attachments or specific interaction.
- Inhibition of adherence of a blood cell or protein to a biological tissue can be measured using methods known in the art. For example, assays for detecting binding of glycoprotein Ib ⁇ to a biological tissue are described in Simon et al., J. Exp. Med. 192:193-204, 2000, and references cited therein. In various embodiments, binding of a GP lb ⁇ fusion protein inhibits binding of a blood cell or protein to a biological tissue by at least 30%, 50%, 75%, 90%, 95%, 99% or 99.9%.
- Adherence can also be assessed in condition of greater or less than physiological flow conditions, including static conditions and serial application of static and shear conditions. Adherence can be determined for example colormetrically, fluorometrically, by flow cytometry or using a parallel plate flow chamber assay.
- the subject is also administered one or more of the following: a statin;acetylsalicylic acid (aspirin);heparin (including unfractionated or low-molecular weight heparins); glycoprotein Ilb/Tfla antagonists; clopidogrel; P-selectin antagonists; thrombin inhibitors; and thromboiytic enzymes.
- the subject can be e.g., any mammal, e.g., a human, a primate, mouse, rat, dog, cat, cow, horse, pig.
- the therapeutics include, e.g. : (i) any one or more of the glycoprotein Ib ⁇ polypeptideand derivative, fragments, analogs and homologs thereof; (ii) antibodies directed against the glycoprotein Ib ⁇ polypeptides described in (i)and (iii) nucleic acids encoding a glycoprotein Ib ⁇ polypeptide, and derivatives, fragments, analogs and homologs thereof as described in (i) above.
- any disorder that is etiologically linked to platelet activation is considered amenable to prevention or to treatment.
- the disorder can be, e.g., vascular inflammation; atherosclerosis; restenosis (e.g., angioplasty-related restenosis); and/or a condition associated with hrombotic disease, e.g., angina, (including stable angina and unstable angina) acute myocardial infarction, stroke, venous thrombosis or arterial thrombosis.
- Example 1 GPIb-Ig tyrosine sulfation plays an important role in thrombin binding but is less important for vWF binding
- the role of the acidic carboxyl terminal region of GPIb ⁇ in binding to vWF and thrombin was examined by tyrosine residues at positions 276, 278, and/or 279 of three types of GPlb ⁇ -IgdFc fusion proteins: those with a wild-type human GPIb ⁇ sequence; a fusion protein with the M239V (IV) substitution, and a fusion protein with a double G233V M239V (2V) substitution.
- the proteins were produced in Chinese Hamster Ovary (CHO) cells, then isolated by Protein A affinity chromatography and anion exchange chromatography. Dimerized forms of the fusion proteins in which the sulfation state varied from 0 to 6 were isolated. Binding of the isoforms to the vWF Al domain (Al), to intact vWF, and to ⁇ -thrombin was examined using surface plasmon resonance (BiaCore).
- the IV and 2V polypeptides bound with significantly higher affinity to the A 1 domain as compared to the WT polypeptides.
- the IV and 2V polypeptides also showed significantly enhanced affinity for intact vWF relative to the affinity of the fusion protein with the wild-type GPIb ⁇ sequence for vWF
- the Ka for the 2V variant ranged from 4 nm to 12 nm
- the Kd for the IV variant ranged from 119 nm to 284 nM for fully sulfated to non-sulfated isoforms. Binding of wild-type GPIb ⁇ to vWF was not detected under the same conditions, which indicates a Kd > 1 mM.
- the binding affinity of the fully sulfated isoform to thrombin was ⁇ 80-fold higher than that of the unsulfated isoform (K D of 0.5 ⁇ M and 40 ⁇ M, respectively).
- K D 0.5 ⁇ M and 40 ⁇ M, respectively.
- K D 3.4 - 15 nM for 2V 6-0 sulfated, 113 - 521 nM for IV 6-0 sulfated.
- the relationship between sulfation state and binding to von Willebrand Factor and binding to thrombin was also examined.
- Rat tail bleeding method Adult male Sprague Dawley rats (Taconic), weighing 250-300gm, were used. The animals were equipped with a jugular vein catheter by the vendor before arriving. The animals were housed in individual cages at room temperature under a light period of 12hr (6:am -6:pm) at our institutional animal facility and were provided standard laboratory rodent chow and water ad libitum. The animals were acclimated to their new environment at least three days before the use.
- GPIb-Ig sample administration and rat Tail vein bleeding procedures On the day of the experiment, an extension tubing (20 cm of PE 50 tubing) with a 3 -way stopcock and a 1-cc syringe filled with sterile isotonic 0.9% saline was attached to the indwelling jugular vein catheter.
- Stock GPIb-Ig test sample was diluted with isotonic 0.9% saline and was administered at indicated doses in a volume of 0.1ml per 100 gm body weight via another arm of the 3-way stopcock. The catheter was flushed with 0.2 ml of saline. Ten minutes after the injection, the rats were removed from their home cage and were gently put into a restrainer.
- the distal 1-2 mm segment of the tail was transected using a razor blade and the tail was quickly immersed into a 300ml-beaker pre-filled with 0.9% isotonic saline (temperature 37°C).
- Bleeding time was defined as the period of time from the start of the tail bleeding to completely cease of bleeding (no re-bleeding within 30 second once the bleeding has stopped).
- the rats were euthanized by inhalation of CO 2 .
- 10 minutes following an i.v. dose of 200ug/kg bleeding time of four minutes was detected in GPIb ⁇ variants with a sulfation state 0, 1, or 2. Bleeding time increased as the sulfation state increased to 6, where bleeding time was 10 minutes.
- the GPIb ⁇ fusion protein variants tested included IV, 2V, 3V, 3V/C65S, 2V/FFF, and a variant named "clipped 2V".
- Control reactions included a 290 amino acid GPIb ⁇ protein, a vWF polypeptide alone or an irrelevant control polypeptide (47.mFc, a PSGL-1 fusion protein).
- a fusion polypeptide or control polypeptide was mixed with Protein A Sepharose beads, which bound to the immunoglobulin moieties of the fusion protein. Protein bound to the Protein A Sepharose beads were eluted and electrophoresed through a 3-8% Tris-Acetate gel. The gel was then immunoblotted with a polyclonal antibody to vWF, and the antibody detected.
- vWF showed the strongest binding to the GPIB ⁇ variants 3V and 3V/C65S when assayed under both reducing and non-reducing conditions. Strong binding was also observed with the 2V variant, while comparatively less binding was detected with the IV variant and the 2V/FFF variant.
- the GPIb ⁇ wild-type polypeptide showed weak binding to vWF in these experiments.
- Example 5 In vivo inhibition of repetitive coronary artery thrombosis
- the ability of a fusion Ib ⁇ protein-immunoglobulin protein to inhibit coronary artery thrombosis in vivo is determined using the procedure described by Folts at al., Circulation 54:365- 70, 1976 is examined.
- the fusion protein includes a GPIb ⁇ sequence with the substitutions G233V K237V and M239V relative to the amino acid sequence of SEQ ID NO: 1.
- Mongrel dogs weighing 20-25kg, are anesthetized with sodium pentobarbital (30mg/kg i.v.), then intubated and ventilated with room air using a respirator.
- Venous and arterial catheters are placed.
- the heart is approached by left thoracotomy through the fifth intercostal space.
- the pericardium is opened and sutured to the wound edges to provide a cradle without displacing the heart.
- About 2cm of the left circumflex coronary artery (LCX) is isolated. Mean and dynamic LCX flow are continuously monitored using a perivascular ultrasonic flow probe placed proximally on the artery.
- the endothelium of the LCX is injured by squeezing with a hemostat.
- a plastic constrictor is placed distal and overlying the area of injured endothelium to provide approximately 70-80% vessel stenosis.
- the blood flow is restored by shaking the constrictor to dislodge aggregated platelets. This decrease and restoration of blood flow are termed CFRs. At least five consecutive CFRs are recorded prior to administering the test drug.
- Example 6 In vivo treatment of a subject with of unstable angina (TJA) or non-ST-elevated mvocardial infarction (NSTEMI)
- a GPIB ⁇ -Ig variant according to the invention is administered as a fusion protein by a single intravenous bolus injection (10 mg) to a patient with unstable angina or non-ST-elevated myocardial infarction (NSTEMI). Treatment with the variant ameliorates the UA or NSTEMI.
- the ristocetin induced platelet aggregation assay (RIP A) is used to monitor GPIb-Ig activity.
- the GPIb ⁇ variant was administered intravenously two hours after administration of clopidogrel (4.3 mg/kg, po).
- GPIb ⁇ variants tested included GPIb ⁇ 2V/290/FFF GPIba 2V/290 GPIba 2V/290 GPIba 2V/290, GPIba 2V/290/FFF (100 mcg/kg), and GPIba 2V/290 (50 mcg/kg).
- RBT was measured 15 minutes after administration of the GPI ⁇ variant. Six or seven animals were tested for each treatment.
- Example 8 Comparison of antithrombotic efficacy of sulfated and non-sulfated forms of recombinant soluble GPIba chimeras in a canine model of coronary artery thrombosis
- GPIb ⁇ -290/2V/FFF-Ig was generated by further substituting three tyrosine residues (Tyr-276, 278, 279) with phenylalanines to eliminate sulfation.
- Cyclic flow reductions were induced by crush injury of the left circumflex coronary artery in anesthetized male mongrel dogs. Coronary blood flow was continuously monitored using a perivascular ultrasonic flow probe. The template bleeding time was measured at the surface of the inner upper lip using an automated cutting device. The bleeding time prolongation caused by sulfated and non-sulfated GPIba chimeras was assessed by using rat tail bleeding model. Platelet function in whole blood before and at various time points after treatment in Folts' model was measured with a Platelet Function Analyzer- 100 (PFA-100).
- PFA-100 Platelet Function Analyzer- 100
- FIGS. 1A-1D The effect of wild-type, or GPIb chimeras on ristocetin-induced human platelet aggregation is shown in FIGS. 1A-1D. Shown are wild-type GPIba ("Normal"), GPIb-Ig WT , GPIba-290/2V- Ig, and GPIba-290/2V/FFF-Ig.
- FIG. 2 is a graph showing the antithrombotic efficacy of GPIb ⁇ chimeras in a canine Folts' model of coronary thrombosis (y-axis: LCX blood)
- FIG. 3 is a graph showing inhibition of platelet function by GPIb/290/2V-Ig and GPIb/290/2 V/FFF-Ig in a canine model of coronary thrombosis assessed by PFA- 100.
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Abstract
La présente invention concerne des compositions et des méthodes de traitement ou de prévention de troubles vasculaires.
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US (2) | US7727535B2 (fr) |
EP (2) | EP2522677A1 (fr) |
JP (1) | JP2007537710A (fr) |
KR (1) | KR20060022261A (fr) |
CN (1) | CN100436480C (fr) |
AU (1) | AU2004247737B2 (fr) |
BR (1) | BRPI0411289A (fr) |
CA (1) | CA2530972A1 (fr) |
HK (1) | HK1095152A1 (fr) |
IL (2) | IL172337A (fr) |
MX (1) | MXPA05013509A (fr) |
NO (1) | NO20060093L (fr) |
NZ (1) | NZ544165A (fr) |
RU (1) | RU2403262C2 (fr) |
WO (1) | WO2004111089A2 (fr) |
ZA (1) | ZA200510331B (fr) |
Families Citing this family (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7534595B2 (en) * | 2006-06-12 | 2009-05-19 | Biomarin Pharmaceutical Inc. | Compositions of prokaryotic phenylalanine ammonia-lyase and methods of using compositions thereof |
US11535673B2 (en) * | 2007-04-05 | 2022-12-27 | President and Fellows of Harvard CoHege | Chimeric activators: quantitatively designed protein therapeutics and uses thereof |
DE102007031708A1 (de) * | 2007-07-06 | 2009-01-08 | Dade Behring Marburg Gmbh | Bestimmung der von Willebrand Faktor-Aktivität in Abwesenheit von Ristocetin |
EP2193373A4 (fr) | 2007-08-23 | 2010-07-28 | Bloodct Of Wisconsin Inc | Procédés et kits pour mesurer le facteur von willebrand |
AU2012339618B2 (en) * | 2011-11-15 | 2017-09-07 | The Walter And Eliza Hall Institute Of Medical Research | Soluble mediator |
WO2013096932A1 (fr) * | 2011-12-23 | 2013-06-27 | Vasculogics, Inc. | Compositions et procédés pour le traitement de thrombose et la prolongation de la survie de plaquettes stockées |
ES2614352T3 (es) * | 2014-03-11 | 2017-05-30 | Siemens Healthcare Diagnostics Products Gmbh | Método para la detección de moduladores de la interacción de GPIb-trombina |
EP3139955B1 (fr) | 2014-04-30 | 2024-03-20 | President and Fellows of Harvard College | Protéines de fusion pour le traitement du cancer et procédés associés |
CN106661119A (zh) | 2014-07-01 | 2017-05-10 | 辉瑞公司 | 双特异性异二聚化双抗体及其用途 |
EP4386387A2 (fr) * | 2018-02-20 | 2024-06-19 | F. Hoffmann-La Roche AG | Composition comprenant une protéine recombinante du récepteur gpiba |
CN113264995A (zh) * | 2021-05-26 | 2021-08-17 | 苏州大学 | 一种血小板GPIbα蛋白相关的抗原表位肽及其应用 |
WO2023007182A1 (fr) * | 2021-07-29 | 2023-02-02 | The University Of Birmingham | Inhibiteurs d'interactions de protéines |
WO2023109903A1 (fr) * | 2021-12-15 | 2023-06-22 | 上海循曜生物科技有限公司 | Produit et méthode de traitement de la thrombocytopénie |
Family Cites Families (17)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5340727A (en) * | 1987-11-17 | 1994-08-23 | The Scripps Research Institute | GPIbα fragments and recombinant DNA expression vectors |
IL88308A0 (en) * | 1987-11-17 | 1989-06-30 | Scripps Clinic Res | Peptides that block the binding of von willebrand factor |
US4899195A (en) * | 1988-01-29 | 1990-02-06 | Ushio Denki | Method of exposing a peripheral part of wafer |
US5225538A (en) | 1989-02-23 | 1993-07-06 | Genentech, Inc. | Lymphocyte homing receptor/immunoglobulin fusion proteins |
US5013556A (en) | 1989-10-20 | 1991-05-07 | Liposome Technology, Inc. | Liposomes with enhanced circulation time |
US5194743A (en) * | 1990-04-06 | 1993-03-16 | Nikon Corporation | Device for positioning circular semiconductor wafers |
JP2874280B2 (ja) * | 1990-05-16 | 1999-03-24 | 株式会社ニコン | 周縁露光装置及び周縁露光方法 |
US6136310A (en) | 1991-07-25 | 2000-10-24 | Idec Pharmaceuticals Corporation | Recombinant anti-CD4 antibodies for human therapy |
US5317097A (en) * | 1991-10-07 | 1994-05-31 | The Research Foundation Of State University Of New York | Mutations in the gene encoding the α chain on platelet glycoprotein IB |
US6277975B1 (en) * | 1992-10-23 | 2001-08-21 | Genetics Institute, Inc. | Fusions of P-selectin ligand protein and polynucleotides encoding same |
US5516964A (en) * | 1994-01-21 | 1996-05-14 | Sun Company, Inc. (R&M) | Hydrocarbon isomerization using solid superacid catalysts comprising platinum metal |
TW316322B (fr) * | 1995-10-02 | 1997-09-21 | Ushio Electric Inc | |
JP3237522B2 (ja) * | 1996-02-05 | 2001-12-10 | ウシオ電機株式会社 | ウエハ周辺露光方法および装置 |
JP3735921B2 (ja) * | 1996-02-07 | 2006-01-18 | 三菱ウェルファーマ株式会社 | GPIb・脂質複合体およびその用途 |
ES2532910T3 (es) | 1998-04-02 | 2015-04-01 | Genentech, Inc. | Variantes de anticuerpos y fragmentos de los mismos |
EP1074564B1 (fr) | 1998-04-23 | 2008-12-10 | Ajinomoto Co., Inc. | Substance possedant une activite antithrombotique et procede de detection de glycokallidine |
AR035779A1 (es) * | 2001-02-06 | 2004-07-14 | Genetics Inst Llc | Polipeptidos de fusion derivados de glicoproteina ib alfa de plaqueta y metodos de uso de los mismos |
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2004
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- 2004-06-14 KR KR1020057023647A patent/KR20060022261A/ko not_active Application Discontinuation
- 2004-06-14 MX MXPA05013509A patent/MXPA05013509A/es active IP Right Grant
- 2004-06-14 WO PCT/US2004/019057 patent/WO2004111089A2/fr active Application Filing
- 2004-06-14 EP EP12174152A patent/EP2522677A1/fr not_active Withdrawn
- 2004-06-14 NZ NZ544165A patent/NZ544165A/en not_active IP Right Cessation
- 2004-06-14 CA CA002530972A patent/CA2530972A1/fr not_active Abandoned
- 2004-06-14 EP EP04776599A patent/EP1636266A2/fr not_active Withdrawn
- 2004-06-14 RU RU2006100306/10A patent/RU2403262C2/ru not_active IP Right Cessation
- 2004-06-14 ZA ZA200510331A patent/ZA200510331B/xx unknown
- 2004-06-14 US US10/868,371 patent/US7727535B2/en not_active Expired - Fee Related
- 2004-06-14 AU AU2004247737A patent/AU2004247737B2/en not_active Ceased
- 2004-06-14 BR BRPI0411289-0A patent/BRPI0411289A/pt not_active IP Right Cessation
- 2004-06-14 JP JP2006533797A patent/JP2007537710A/ja active Pending
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2005
- 2005-12-02 IL IL172337A patent/IL172337A/en not_active IP Right Cessation
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2006
- 2006-01-06 NO NO20060093A patent/NO20060093L/no not_active Application Discontinuation
-
2007
- 2007-02-22 HK HK07102003.4A patent/HK1095152A1/xx not_active IP Right Cessation
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2009
- 2009-11-19 US US12/621,887 patent/US20110039780A1/en not_active Abandoned
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2012
- 2012-06-10 IL IL220272A patent/IL220272A0/en unknown
Non-Patent Citations (1)
Title |
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See references of WO2004111089A2 * |
Also Published As
Publication number | Publication date |
---|---|
AU2004247737B2 (en) | 2009-04-23 |
US7727535B2 (en) | 2010-06-01 |
RU2006100306A (ru) | 2006-06-10 |
NZ544165A (en) | 2008-12-24 |
ZA200510331B (en) | 2007-03-28 |
NO20060093L (no) | 2006-03-01 |
IL172337A (en) | 2012-07-31 |
WO2004111089A2 (fr) | 2004-12-23 |
CN100436480C (zh) | 2008-11-26 |
IL220272A0 (en) | 2012-07-31 |
HK1095152A1 (en) | 2007-04-27 |
RU2403262C2 (ru) | 2010-11-10 |
AU2004247737A1 (en) | 2004-12-23 |
CA2530972A1 (fr) | 2004-12-23 |
KR20060022261A (ko) | 2006-03-09 |
BRPI0411289A (pt) | 2006-08-01 |
EP2522677A1 (fr) | 2012-11-14 |
MXPA05013509A (es) | 2006-04-05 |
CN1823090A (zh) | 2006-08-23 |
US20050089888A1 (en) | 2005-04-28 |
US20110039780A1 (en) | 2011-02-17 |
WO2004111089A3 (fr) | 2005-05-12 |
JP2007537710A (ja) | 2007-12-27 |
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